HDAC6 Inhibitors Rescued the Defective Axonal Mitochondrial Movement in Motor Neurons Derived from the Induced Pluripotent Stem Cells of Peripheral Neuropathy Patients with HSPB1 Mutation

Figure 3

Axonal transport of mitochondria in control MNs, S135F-MNs and P182L-MNs. (a) Microfluidic culturing system. Microchannel plates were made from transparent polymer in a specialized mold to create multicompartments connected by μm-sized grooves (833.4 μm long in length) that only allow growing axons to pass through, thereby separating the microenvironments of neuronal components. (b) Visualization of mitochondria by Mito-dsRED2 transfection (upper image) and kymograph images for mitochondria movement (middle and lower four images). Mito-dsRED2 vector was introduced into fully differentiated MNs cultured on microchannel plates for an additional 1 week. Mitochondrial images were taken at a ratio of 121 snaps/2 min and stacked into kymograph for analysis (original magnification, 1000x). (c) S135F-MNs show decreased velocity of moving mitochondria compared to control MNs. Unpaired t-tests,, , and . Absolute velocity of moving mitochondria was calculated from measured angle and length in kymograph images using ImageJ (WA09-MNs; , hFSiPS1-MNs; , S135F-MNs; , and P182L-MNs; ). (d) S135F-MNs and P182L-MNs show reduced moving proportion of mitochondria in an axon (WA09-MNs; , hFSiPS1-MNs; , S135F-MNs; , and P182L-MNs; ).

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